Engineering transcription factor BmoR for screening butanol overproducers.

Amino Acid Substitution Bacterial Proteins / chemistry Binding Sites Butanols / chemistry Mutagenesis, Site-Directed Mutation, Missense Transcription Factors / chemistry
BmoR Detection range Industrial significance Protein engineering Screening Transcription factor

Journal

Metabolic engineering
ISSN: 1096-7184
Titre abrégé: Metab Eng
Pays: Belgium
ID NLM: 9815657

Informations de publication

Date de publication:
12 2019
Historique:
received: 05 05 2019
revised: 09 07 2019
accepted: 22 08 2019
pubmed: 27 8 2019
medline: 23 4 2020
entrez: 27 8 2019
Statut: ppublish

Résumé

The wild-type transcription factors are sensitive to their corresponding signal molecules. Using wild-type transcription factors as biosensors to screen industrial overproducers are generally impractical because of their narrow detection ranges. This study took transcription factor BmoR as an example and aimed to expand the detection range of BmoR for screening alcohols overproducers. Firstly, a BmoR mutation library was established, and the mutations distributed randomly in all predicted functional domains of BmoR. Structure of BmoR-isobutanol complex were modelled, and isobutanol binding sites were confirmed by site-directed mutagenesis. Subsequently, the effects of the mutations on the detection range or output were confirmed in the BmoR mutants. Four combinatorial mutants containing one increased-detection-range mutation and one enhanced-output mutation were constructed. Compared with wild-type BmoR, F276A/E627N BmoR and D333N/E627N BmoR have wider detection ranges (0-100 mM) and relatively high outputs to the isobutanol added quantitatively or produced intracellularly, demonstrating they have potential for screening isobutanol overproduction strains. This work presented an example of engineering the wild-type transcription factors with physiological significance for industrial utilization.

Identifiants

DOI: 10.1016/j.ymben.2019.08.015 PMID: 31449878
pubmed: 31449878
pii: S1096-7176(19)30199-5
doi: 10.1016/j.ymben.2019.08.015
pii:
doi:

Substances chimiques

Bacterial Proteins 0
Butanols 0
Transcription Factors 0
isobutyl alcohol 56F9Z98TEM

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

28-38

Informations de copyright

Copyright © 2019 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Auteurs

Huan Yu (H)

Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, 100081, Beijing, China.

Zhenya Chen (Z)

Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, 100081, Beijing, China.

Ning Wang (N)

Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, 100081, Beijing, China; UCLA Institute for Technology Advancement (Suzhou), 10 Yueliangwan Road, Suzhou Industrial Park, 215123, Suzhou, China.

Shengzhu Yu (S)

Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, 100081, Beijing, China.

Yajun Yan (Y)

School of Chemical, Materials and Biomedical Engineering, College of Engineering, The University of Georgia, Athens, 30602, GA, USA.

Yi-Xin Huo (YX)

Key Laboratory of Molecular Medicine and Biotherapy, School of Life Science, Beijing Institute of Technology, No. 5 South Zhongguancun Street, 100081, Beijing, China; UCLA Institute for Technology Advancement (Suzhou), 10 Yueliangwan Road, Suzhou Industrial Park, 215123, Suzhou, China. Electronic address: huoyixin@bit.edu.cn.

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Classifications MeSH

questionsmedicales.fr › Phénomènes génétiques › Mutagenèse › Substitution d'acide aminé › Engineering transcription factor BmoR for...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Engineering transcription factor BmoR for...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Sites de fixation › Engineering transcription factor BmoR for...
questionsmedicales.fr › Lipides › Alcools gras › Butanols › Engineering transcription factor BmoR for...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Ingénierie des protéines › Mutagenèse dirigée › Engineering transcription factor BmoR for...
questionsmedicales.fr › Phénomènes génétiques › Variation génétique › Mutation › Mutation faux-sens › Engineering transcription factor BmoR for...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Facteurs de transcription › Engineering transcription factor BmoR for...